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Method for establishing rat contrast-induced nephropathy model

A technology of contrast agent nephropathy and a construction method, which is applied in the field of construction of a rat contrast agent nephropathy model, can solve the problems of not significantly improving the success rate of modeling, unable to obtain blood volume changes, increasing the mortality rate of model animals, etc. Success rate and effect of simplified steps and reduced vascular condition

Inactive Publication Date: 2019-05-21
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Healthy rats have good tolerance to contrast medium, but the renal injury caused by contrast medium alone has poor reproducibility
Water deprivation may promote the effective renal injury of contrast media by prolonging the metabolic cycle of contrast media in the kidney; however, water deprivation leads to a decrease in renal blood flow and a longer contact time with contrast media, and previous methods of exploring water deprivation for several hours could not obtain obvious effects. Changes in blood volume play a synergistic effect, so the time of water deprivation is an important factor affecting the success rate of the model
[0004] Furosemide is a diuretic, which was injected through the tail vein 2 hours before the contrast medium in previous methodological reports; however, from the exploration of the inventors of this application, it was found that furosemide injection not only did not significantly improve the success rate of modeling , on the contrary, it will affect the vascular conditions when the contrast agent is injected through the tail vein and increase the mortality of model animals

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  • Method for establishing rat contrast-induced nephropathy model
  • Method for establishing rat contrast-induced nephropathy model
  • Method for establishing rat contrast-induced nephropathy model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The impact of embodiment 1 water deprivation time on contrast agent nephropathy model

[0028] Test method: as figure 1 As shown, SD rats aged 7-8 weeks, weighing 180-220 g, were collected by retro-orbital vein blood collection method before operation to collect basic venous blood samples to detect related indicators of renal function. 5 rats in each group, randomly divided into (A) water deprivation group 4hrs, (B) 24hrs, (C) 48hrs and (D) 72hrs and (E) non-water deprivation furosemide group (tail vein injection of furosemide stock solution 10ml / kg ) a total of 5 groups. After the experimental operation, inject 10ml / kg of normal saline (32G needle) through the tail vein and open drinking water. After 24hrs, blood was collected and centrifuged at 2500rpm for 15min to extract the upper clear serum. The changes of serum creatinine (Scr), cystatin C (Cys-C) and blood urea nitrogen (BUN) were detected by immunoluminescence method.

[0029] Test result: if figure 2 As sh...

Embodiment 2

[0033] Example 2 Construction method of rat contrast agent nephropathy model and serological detection of changes in rat renal function

[0034] In order to explore how to better model successfully in combination with intravenous injection of contrast agents under fixed water-deprivation conditions, the following experimental methods were adopted:

[0035] see image 3 , SD rats aged 7-8 weeks, weighing 180-220g, were collected from the retro-orbital vein before the operation, and basic venous blood samples were collected to detect related indicators of renal function. There were 6 rats in each group, and the water deprivation time was set at 48hrs. The experimental groups were a: furosemide + NS (normal saline) control group; b: furosemide + CM (contrast medium) group; c: water deprivation + NS group; d: water deprivation + furosemide group; e: water deprivation + CM group; f: water deprivation + furosemide + CM group. Blood was collected 24hrs and 48hrs after modeling to ...

Embodiment 3

[0043] Example 3 Observation of kidney tissue morphology by HE staining reaction

[0044] Experimental method: Whole kidney tissues of the CIN model group and NS control group (treated with normal saline) were cross-sectioned 5-10 μm and stained with HE. The dyeing method is a standard operation. Stained slides were observed under light microscope for changes in renal cortex, cortex-medull junction and medulla area.

[0045] Experimental results: if Figure 5 As shown, in the CIN model group (consistent with the modeling method of group e in Example 2) the renal histological changes of the rats are obvious, and the heavily damaged parts are concentrated in the proximal tubules of the cortical-medullary junction and the outer intramedullary zone, mainly It manifests as vacuolar degeneration of tubular epithelial cells, flattened atrophy or necrotic shedding of cells, and part of the basement membrane is exposed; dilation of the lumen can be seen, and protein casts are occasio...

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Abstract

The invention discloses a method for establishing a rat contrast-induced nephropathy model. The method includes the following steps that 1, rats do not drink water within 24-55 hours; 2, iopromide isapplied to the rats who do not drink water within 24-55 hours to obtain the contrast-induced nephropathy model. After detecting renal damage markers such as serum creatinine after different water-forbidden time in the test, it comes to a conclusion through discussion that occurrence of CIN is promoted under suitable water-forbidden conditions, and there are no irreversible renal damage changes caused by water-forbidden operation factors. The operation steps in the modeling process are optimized, caudal vein furosemide injection operation is reduced, good vascular conditions are kept for injection of contrast agent, unnecessary intervention factors are reduced, the success rate of tail vein injection of the contrast agent and the success rate of model construction are increased, and the simplified and repeatable modeling process can be completed with fewer and effective steps.

Description

technical field [0001] The invention relates to the technical field of constructing a nephropathy animal model, in particular to a method for constructing a rat contrast medium nephropathy model. Background technique [0002] Acute contrast-induced renal injury (CIN) is an acute kidney injury syndrome that occurs 24-72 hours after exposure to iodine-containing contrast media, manifested by elevated serum creatinine (SCr), transient proteinuria, and oliguria. Acute contrast-induced kidney injury, also known as contrast-induced nephropathy, will increase renal replacement therapy and mortality in clinical patients. The current research on the mechanism of contrast-induced nephropathy believes that it is a comprehensive effect, mainly including direct renal tubular toxicity of contrast medium, renal interstitial ischemia and oxidative stress. At present, the treatment measures can only focus on prevention, and there is no standardized and effective treatment plan. Therefore, e...

Claims

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Application Information

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IPC IPC(8): A61K31/166A01K67/027
Inventor 崔同涛罗碧辉王慧勇王涛陆东风
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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